LOCUS       sequence-390280-        5498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_001497284 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5498)
  AUTHORS   Miura F, Miura M, Shibata Y, Furuta Y, Miyamura K, Ino Y, Bayoumi 
            AMA, Oba U, Ito T
  TITLE     Identification, expression, and purification of DNA cytosine 
            5-methyltransferases with short recognition sequences.
  JOURNAL   BMC Biotechnol. 2022 Nov 4;22(1):33. doi: 
            10.1186/s12896-022-00765-3.
  PUBMED    36333700
REFERENCE   2  (bases 1 to 5498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2022-11-4"; volume: "22(1):33. doi"; pages: " 
            10.1186/s12896-022-00765-3"
COMMENT     Sequence Label: pBZS-YP_001497284
FEATURES             Location/Qualifiers
     source          1..5498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1837..1860
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1899..1916)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2069..2155
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2247..2274
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2293..2384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2385..2453,2454..3245)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2603..2622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3416..4004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3905..3924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4158..4175
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4190..4330
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4256..4275)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4416..4438
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4594..5472)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHFFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagccg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa gctcgaggga
      421 tccatgtatt gtatgttgag gtcattagat ctattcagcg gcatcggtgg taactcctat
      481 gctttacgcg acatcctgaa accggttgcg tatgtggagc gcgagcagca tctgcgcgat
      541 ttcctgggtc ggaagtttcc ggacgtcccg attttcgacg acgttgtcac ctttgatacc
      601 agatccgtga aggacatcga tattatcacc gcgggcttcc cgtgcaccgg tttttccacg
      661 gctggcaagg gcgatggatt cgagcacgag gcgagcggtc tgtttaccga ggtggtgcgc
      721 atcgccaaag aactggaacc gcgttttgtt tttttggaga actcacatac cgttgcccgt
      781 gtcgagaacc tgcacgtgat tatcgacgcc ttcgatgtgc tgggctacga ctgtcgttgg
      841 acgaccaccc atgcaactgc ggtgggtgcg cctcaacaac gccatcgttg gttctgcttg
      901 gctgtgcgtc gtggtgaaag caccgatgtg gacatcccgg acgttgacca ctttgattgg
      961 accaatgatg aaccagcgaa gcagattgaa aacattgaca gccgtggtcg taagatcatc
     1021 caggcgctcg gcaactctat cgttccggac cagctgagaa gcgcattcaa gaccattatg
     1081 agcatggatt ttgacggcaa tgaaacccag ggtagcaaac gcatcccgca cggctactcg
     1141 atccgcggta aaatcttcaa gaagaacatc attaacaccg agcgtgaacc gatgaatatc
     1201 ctgatttgcc aatctgaacc accgcaaaaa cacaaaggcc gtctgccgca cattgataaa
     1261 aagattaaac gtttctgggc aactccggtt aagggcacgg cgacgaaagg tcagaccatt
     1321 ttgacccgtc gcagcctgca atgtctgggt acattggttc gtttctctcc ggatggtatt
     1381 agcggctggc atttgaatcc gttttgggtt ggctacctga tgggttatcg tcgtgacttc
     1441 ttcgaggaat ttgaattcca gcatgatgaa gcagtggata acaagtttaa caaagaacag
     1501 cagaacgcgt tttatgagat cttgcactta ccgaatctga acgaggaaca gcgcaatgcg
     1561 ttcattcagt cccttaaaga cgatccatct caaagcgcaa atctgctggc agaagccaag
     1621 aaactgaacg atgctcaagc tccgaaagtc gacaacaaat tcaacaaaga gcaacagaat
     1681 gcgttctacg aaattctgca tttaccgaac ctgaatgaag aacagcgtaa cgcctttatc
     1741 caatcgctga aagatgatcc tagtcagtca gccaatctgt tggcggaagc gaagaaactc
     1801 aatgacgccc aagctcccaa agttgacgcg aatagctgga gccatccgca gtttgaaaag
     1861 ctttagggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     1921 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     1981 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     2041 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2101 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     2161 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     2221 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     2281 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     2641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3241 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3541 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4081 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4141 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4201 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     4261 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4321 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4381 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4441 ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg
     4501 gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     4561 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
     4621 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
     4681 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
     4741 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
     4801 taagacgcta atccctaact gctggcggaa aaaatgtgac agacgcgacg gcgacaagca
     4861 aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta
     4921 ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc
     4981 catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc
     5041 ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc
     5101 ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg
     5161 ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg
     5221 acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac
     5281 aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata
     5341 acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg
     5401 cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg
     5461 cgcttcagcc atacttttca tactcccgcc attcagag
//
